Smooth to Rugose phase variation in Vibrio cholerae

霍乱弧菌导致霍乱，是水生动物的自然居住者 环境。季节性霍乱暴发发生在疾病流行并可传播的地方 全世界。S霍乱弧菌引起疫情的能力与其在水生环境中的生存能力有关 栖息地。已经提出了霍乱弧菌？S形成生物膜的能力，基质包裹， 与表面相关的群落，对其在疫情期间在水生生境中的生存至关重要 并且在流行期间有利于宿主间的传播。这样做的目的是 建议是为了提高我们对生物膜基质成分、机理和 生物膜形成的调控，环二聚鸟苷一磷酸(CdiGMP)的作用机制 信号，以及它们在霍乱弧菌生物学中的重要性。在目标1中，我们将重点放在 生物膜基质成分的表征。我们将确定弧菌的结构 多糖(VPS)和VPS生物合成所需的VPS簇基因和 生物膜的形成。我们将确定VPS结合能力和定位模式 生物膜中的基质蛋白，并检验我们的假设，即这些蛋白与VPS的糖结合 以稳定母体。我们还将研究推测的一种酶的性质 VPS裂解酶。最后，我们将确定已知的生物膜决定因素在V。 霍乱致病机制。在目标2中，我们将研究生物膜形成的规律。我们会 检查生物膜基质产生和溶解所需基因的调控特性； 并确定正转录调控因子和负转录调控因子与这些基因的相互作用 推动者。我们还将调查影响监管的环境条件 生物膜基质生产。在目标3中，我们将阐明cdiGMP的分子机制 信号转导控制生物膜的形成。我们将确定cdiGMP的细胞定位。 调节生物膜形成的信号蛋白，以及这些蛋白是否 在牢房里被隔开了。为了鉴定c-di-GMP信号系统的靶蛋白， 我们将寻找与c-diGMP信号蛋白相互作用的蛋白质，以及c-diGMP。 皱纹变异体中的受体蛋白。然后我们将评估这些c-diGMP信号是如何 利用小鼠感染模型，蛋白质影响霍乱弧菌的致病机制。更好地理解 生物被膜的形成机制，c-diGMP信号转导，以及两者的重要性 霍乱弧菌生物学过程将被证明对未来制定霍乱弧菌的战略是有用的 预测和控制霍乱流行，并促进确定新的药物靶点 用于在感染过程中对抗病原体。

Vibrio cholerae causes the disease cholera and is a natural inhabitant of aquatic environments. Seasonal cholera outbreaks occur where the disease is endemic and can spread worldwide. V. cholerae¿s ability to cause epidemics is tied to its ability to survive in aquatic habitats. It has been proposed that V. cholerae¿s ability to form biofilms, matrix-enclosed, surface-associated communities, is crucial for its survival in aquatic habitats between epidemics and is advantageous for host-to-host transmission during epidemics. The objective of this proposal is to improve our understanding of biofilm matrix components, the mechanisms and regulation of biofilm formation, the mechanism of cyclic dimeric guanosine monophosphate (cdiGMP) signaling, and their importance in the biology of V. cholerae. In Aim 1, we will focus on characterization of biofilm matrix components. We will determine the structure of Vibrio polysaccharide (VPS) and the genes of the vps cluster required for VPS biosynthesis and biofilm formation. We will determine VPS binding capacities and localization patterns of the matrix proteins in biofilms, and test our hypothesis that these proteins bind to the sugars of VPS in order to stabilize the matrix. We will also investigate the enzymatic properties of a putative VPS lyase. Finally, we will ascertain the contribution of known biofilm determinants in V. cholerae pathogenesis. In Aim 2, we will investigate the regulation of biofilm formation. We will examine the regulatory features of genes required for biofilm matrix production and dissolution, and determine the interactions of the positive and negative transcriptional regulators with these promoters. We will also investigate the environmental conditions that influence the regulation of biofilm matrix production. In Aim 3, we will elucidate the molecular mechanisms by which cdiGMP signaling controls biofilm formation. We will determine the cellular localization of cdiGMP signaling proteins that modulate biofilm formation, and whether these proteins are compartmentalized in the cell. To identify the target proteins of the c-di-GMP signaling systems, we will search for proteins interacting with c-diGMP signaling proteins, as well as for c-diGMP receptor proteins within the rugose variants. We will then assess how these c-diGMP signaling proteins affect V. cholerae pathogenesis, using a murine infection model. Better understanding of the mechanism of biofilm formation, c-diGMP signaling, and the importance of both of these processes In V. cholerae biology will prove useful for the development of future strategies for predicting and controlling cholera epidemics, and to facilitate identification of novel drug targets for combating the pathogen during infection.